AIM: To investigate the effects of propofol on the expression of tissue-type plasminogen activator (tPA) and matrix metalloproteinase 9 (MMP9) in the hippocampus and the cognitive function in neonatal rats.METHODS: The 7-day-old rats were randomly divided into 3 groups: the rats in control (CON) group were intraperitoneally injected with normal saline for 7 d;the rats in single dose of propofol anesthesia (SP) group were intraperitoneally injected with normal saline for 6 d and with propofol on the 7th day;the rats in repeated dose of propofol anesthesia (RP) group were intraperitoneally injected with propofol for 7 d.Blood glucose and blood gas analysis were tested in 6 rats of each group.The rats were randomly selected from each group to isolate the hippocampal tissues at 2 h, 24 h, 48 h, 72 h and 30 d after the last injection.The spatial learning and memory functions of the other rats aged 25 d were determined by Morris water maze.The morphological changes of the hippocampus were observed by HE staining and Nissl's staining.The expression of tPA and MMP9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS: Compared with group CON, the protein expression of tPA and MMP9 in RP group was significantly decreased at each time point, while no significant decrease was observed in SP group except at the time point of 24 h.Compared with CON group, the mRNA expression of tPA and MMP9 was down-regulated obviously in RP group, which was not significantly down-regulated in SP group.From the 3rd training day of Morris water maze beginning, the escape latency was prolonged, and the space exploration time and the number of crossing the original platform location were reduced in RP group compared with CON group and SP group, while no significant difference was observed between CON group and SP group.Compared with CON group, the number of nerve cells reduced and nerve cells arranged in disorder in the hippocampus in RP group.Moreover, the number of Nissl body decreased significantly and finally developed into neuronal degeneration and necrosis in RP group, and no significant difference between SP group and CON group was observed.CONCLUSION: Repeated dose of propofol anesthesia leads to long-term cognitive dysfunction in neonatal rats, which may be related to the down-regulation of tPA and MMP9 expression and destruction of normal morphology and function of neurons in hippocampus, whereas single dose of propofol anesthesia has no such effects.

Animals ; Bufo bufo ; Caspase 3 ; genetics ; metabolism ; Female ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Objective To improve the current standard method of measuring urinary iodine by As (Ⅲ)-Ce4+catalytic spectrophotometry, reducing the amount of arsenic toxic reagent used to decrease environmental pollution,and make the modified method with good precision and accuracy. Methods For improving the current standard method of measuring urinary iodine, amount of arsenious acid solution was reduced from 0.100 mol/L H3AsO3(which contains NaCl 25 g/L) 2.5 ml to 0.025 mol/L H3AsO3(which contains NaCl 40 g/L) 2.5 ml;amount of ceric ammonium sulfate solution was reduced from 0.076 mol/L 0.30 ml to 0.025 mol/L 0.30 ml;photometric wavelength was changed from 420 nm to 400 nm. The new modified method was evaluated by standard curve linearity and linear range, sample detection limit, precision, accuracy, and urinary iodine values, and the rates of absorbance change in the test process were compared with the current standard method. Results The calibration relation of C= a + blgA (C: iodine concentration, A : measuring absorbance) in the new modified method existed when As3+-Ce4+ catalytic reaction was kept at a certain stable temperature range between 20 - 35 ℃ and in certain stable reacting time. The linear range of the calibration curve was 0 - 300 μg/L and the linear correlative coefficient was - 0.9998. The detection limit for iodine was 4 μg/L(0.25 ml of urine was tested). The test coefficient of variations(CV) were 1.7%(1.1/66.0), 1.8%(1.4/76.0), 2.0%(3.0/147.5), 1.6%(4.2/265.5) when measuring urine samples with iodine concentration of 66.0, 76.0, 147.5, 265.5 μg/L, respectively. The average recovery was 100.6% with a range of 95.0% (57.0/60.0) - 103.7% (62.2/60.0) when measuring 4 urine samples containing different iodine concentration, and average recovery was 101.0% (40.4/40.0), 100.4% (100.4/I00.0), 100.5% (60.3/60.0),100.4% (100.4/100.0), respectively. The test results of two national standard urinary iodine were all within the given value range and the relative deviation(RD) was < 5.0% and < 2.0% in 20, 25, 30, 35 ℃ test temperature,respectively. No significant difference was found between the results of the 48 urine samples determined by the new modified method and the current standard method(t = 0.634, P > 0.05). The table of suitable combination of As3+-Ce4+ reaction temperature and time for this method was obtained(such as 20 ℃ and 53 min, 25 ℃ and 40 min, 30 ℃and 30 min, etc. ). Compared with the standard method, the rates of absorbanee change of As ( Ⅲ )-Ce4+ reaction in the new modified method were more slowly, which further reducing the determination deviation caused by the temperature fluctuations or measuring time deviation in measurement process. Conclusions This new modified method greatly reduces the amount of arsenic in waste, reduces pollution, saving reagents, and this method is easier to operate with better precision and accuracy, which is suitable for application of measuring iodine in urine.

Aim To investigate the role of matrix metalloproteinase-9 down-regulation in the learning and memory dysfunction induced by propofol treatment in rats.Methods 7-day-old SD rats were randomly divided into three groups(n=18):control group(NS group) and repeated doses of propofol group(RP group) was intraperitoneally injected with normal saline and propofol respectively for consecutive seven days, single dose of propofol group(SP group) were intraperitoneally injected with normal saline first for consecutive six days, and then injected with propofol on 7th day.The blood gas and glucose levels were monitored of six rats randomly selected from each group.Morris water maze was conducted to test the learning and memory functions of the remaining rats.The expression of MMP-9, BDNF and caspase-3 was detected by Western blot, and the hippocampal neuron apoptosis was determinated by TUNEL staining.Results Compared with NS group and SP group, the escape latency in RP group was prolonged significantly, exploration time and the number of crossing the platform in RP group were markedly decreased(P<0.05).The expressions of MMP-9 and mBDNF in RP group declined, but the expression of proBDNF and the ratio of proBDNF/mBDNF in RP group were higher than those in NS group and SP group(P<0.05).Compared with NS group and SP group, the number of apoptotic neurons and the expression of cleaved-caspase-3 in RP group were increased significantly, but the expression of pro-Caspase3 in RP group was reduced(P<0.05).There was no difference between SP group and NS group regarding all the results(P>0.05).Conclusions Repeated exposure to propofol can lead to a decline in long-term learning and memory functions in neonatal rats, which may be related to the down-regulation of MMP-9 expression, proBDNF and mBDNF conversion disorder in hippocampus and the apoptosis of hippocampal neurons.However, single exposure to propofol has no significant effect.

ObjectiveTo observe the effect of reduced glutathione(GSH) on expression of malondialdehyde(MDA),glutathione peroxidase(GSH-PX) and superoxide dismutase(SOD) after focal cerebral infarction in rats.MethodsRat models of middle cerebral artery occlusion(MCAO) were estabilished with thread after 2-hour ischemia and 6-hour reperfusion.Rats were divided at random into three groups,i.e.,sham-operated,control and treatment group(with GSH 1200 mg/kg) respectively.After the rats model was performed,neurology deficit score,the size of brain infarct region and the change of brain tissue pathologic were evaluated.Contents of MDA and activity of SOD and GSH-PX were detected with spectrophotometer.ResultsCompared with the control group,GSH can ameliorate neurological deficit score and decrease infarct volume induced by MCAO.GSH may reduce contents of MDA and improve activity of SOD and GSH-PX in brain tissue.ConclusionGSH may reduce contents of MDA and improve activity of SOD and GSH-PX so as to enhance capability of eliminating oxygen free radical,and play a neuroprotective effect after cerebral focal ischemia reperfusion.

Objective To study the dynamic changes in cardiac morphology and myocardial collagen in apolipoprotein E-deficient (apoE-/-) mice of different age with cholesterol-fed. Methods 18 male apoE-/- mice were randomized into 3 groups: 24 weeks group （n=6）, 32 weeks group （n=6）, 40 weeks group （n=6）. 8 C57BL/6J mice were set as the control. Serum cholesterol, low-density lipoprotein cholesterol (LDL-C), triglyceride, high-density lipoprotein cholesterol (HDL-C) were determined. The cardiac morphological changes and the myocardial collagen were observed. Results Compared with the 24 weeks group, the content of myocardial collagen of 32 and 40 weeks group significantly increased (P<0.05). Conclusion With the increasing of age, TC significantly increased, myocardial collagen content tended to go up.

Objective:To observe the effect of liver-soothing and mind-regulating acupuncture method on the resting-state electroencephalography (EEG) in rats with post-traumatic stress disorder (PTSD),and to provide evidence for the effect mechanism study and clinical application of acupuncture intervention for PTSD.Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group,a model group,a grasping group,a paroxetine group and an acupuncture group,with 12 rats in each group.Except for rats in the blank control group,rats in the other groups were subjected to preparing the PTSD models using 'incarceration plus electric shock' method.After interventions,changes in rat behavior of each group were observed;changes in resting-state EEG were collected and analyzed with multichannel EEG acquisition and analysis system,and image analysis and statistical processing were performed.Results:Compared with the blank control group,the average escape latency in the model group was significantly longer (P＜0.05),and the times of crossing the platform and the effective areas were all significantly reduced (P＜0.01).Compared with the grasping group,the average escape latencies in the paroxetine group and acupuncture group were significantly shortened (P＜0.05),and the times of crossing the platform and the effective areas were all significantly increased (P＜0.05).There were no significant differences in the average escape latency,the times of crossing the platform and the effective areas between the acupuncture group and paroxetine group (all P＞0.05).Compared with the blank control group,the α-wave power spectrum value in the model group was significantly decreased,and the power spectrum values of β-wave,δ-wave and a-wave were significantly increased (all P＜0.01);compared with the grasping group,α-wave power spectrum values in the paroxetine group and acupuncture group were significantly increased (both P＜0.01),and the power spectrum values of β-wave,δ-wave and a-wave were decreased significantly (all P＜0.01).The power spectrum values of α-wave,β-wave,δ-wave and (e)-wave of rats in the acupuncture group were not significantly different from those in the paroxetine group (all P＞0.05).Conclusion:Liver-soothing and mind-regulating acupuncture method can significantly improve the abnormal EEG activity in PTSD rats,which may be one mechanism of liver-soothing and mind-regulating acupuncture method in effectively affecting the brain function in PTSD rats.

ObjectiveTo establish a new method with low usage amount of arsenic trioxide for measuring 300 - 1200 μg/L high concentration iodine in urine by As3+-Ce4+ catalytic spectrophotometry using ammonium persulfate digestion, which would be convenient for monitoring urinary iodine in excessive iodine regions and to reduce environmental arsenic pollution. Methods Calibrators and urine samples(0.20 ml each) were digested according to the current standard detection method of urinary iodine(WS/T 107-2006). At the same time, improving the current standard method, the amount of arsenious acid solution was reduced from 0.100 moL/L H3AsO3 (containing NaCl 25 g/L) 2.5 ml to 0.025 mol/L H3AsO3(containing NaCl 40 g/L) 2.5 ml; amount of ceric ammonium sulfate solution was reduced from 0.076 mol/L 0.30 ml to 0.025 mol/L 0.50 ml; and photometric wavelength was changed from 420 nm to 380 nm. The new method was evaluated by standard curve linearity and linear range, sample detection precision, accuracy, and the results of urinary iodine were compared with those determined bycurrent standard method, and this new method was also tested of suitable combination of reaction temperature and reaction time of cerium arsenic in the temperature range of 20 - 30 ℃. Results The calibration relation of C =a + blgA (C: iodine concentration, A : measuring absorhance) in the new method existed when As3+- Ce4+ catalytic reaction was kept at a certain stable temperature range between 20 - 30 ℃ and in certain fixed reacting time. The linear range of the calibration curve was 300 - 1200 μg/L and the linear correlative coefficient was- 0.9999. The relative standard deviations(RSD) were 1.0％(3.2/330.3), 0.4％(2.0/517.3), 0.5％(3.9/712.6) and 0.9％(9.4/1042.3) when measuring urine samples with iodine concentration of 330.3, 517.3,712.6, and 1042.3 μg/L, respectively. The total average recovery was 98.3％ with a range of 93.4％ (186.8/200.0) - 101.5％ (202.9/200.0) when measuring 4 urine samples containing different concentration of high iodine, and average recovery was 99.1％ (148.6/150.0), 97.5％ (195.0/200.0), 98.8％ (395.3/400.0), and 98.2％ (392.7/400.0),respectively. The test results of four urinary iodine standard materials were all within the given value range and the relative deviations(RD) were all ＜ 2.0％ at different test temperature, respectively. No significant difference was found between the results of the 16 urine samples containing high concentration of iodine determined by the new method and the current standard method (|t| =0.727, P ＞ 0.05). The table of suitable combination of As3+-Ce4+ reaction temperature and reaction time for this method was obtained(such as 20 ℃ and 33 min, 25 ℃ and 25 min,30 ℃ and 19 min, etc). Conclusions This method greatly reduces the amount of arsenic in waste, reduces pollution, saves reagents, and this method is easier to be performed with better precision and accuracy, which is suitable for measuring high concentration of iodine in urine.

Child ; Humans ; Lymphoma, T-Cell ; complications ; Panniculitis ; etiology ; Subcutaneous Tissue

OBJECTIVETo establish an HPLC method for the simultaneous determination of three major bioactive components in Jingzhi Guizhi Fuling capsules namely laetrile, paeoniflorin and paeonol.

METHODA LiChrospher C18 column (4.6 mm x 250 mm, 5 microm) was used. The chromatography was carried out with a stepwise gradient programming. The mobile phase was acetonitrile-water (containing 0.1% phosphorous acid) and the flow rate was 1.0 mL x min.

RESULTThe linear range of laetrile was 12.87-102.94 micron x mL(-1), r = 0.999 9, paeoniflorin 24.84 - 198.7 microg x mL(-1), r = 0.9999 and paeonol 12.57-100.56 microg x mL(-1), r = 0.999 9. The method is accurate with variation less than 1.5 % and recovery more than 95 %.

CONCLUSIONThe method was successfully applied to analyze three major bioactive components in Jingzhi Guizhi Fuling capsules.

Acetophenones ; analysis ; Amygdalin ; analysis ; Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Capsules ; Chromatography, High Pressure Liquid ; methods ; Cinnamomum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glucosides ; analysis ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Polyporales ; chemistry ; Reproducibility of Results